蛋白磷酸化与两组分信号系统

可逆的蛋白磷酸化是生物体内存在的一种最为普遍的调节方式,几乎涉及所有的生理及病理过程,在细胞的信号传递中起着极其重要的作用。根据底物蛋白质被磷酸化的氨基酸种类可将各种蛋白激酶分为3类:第一类为丝氨酸/苏氨酸蛋白激酶(ｓｅｒｉｎｅｔｈｒｅｏｎｉｎｅｐｒｏｔｅｉｎｋｉｎａｓｅ,ＳＰＫ),目前发现的蛋白激酶多属此类;第二类为酪氨酸蛋白激酶(ｔｙｒｏｓｉｎｅｐｒｏｔｅｉｎｋｉｎａｓｅ,ＴＰＫ),这类激...     

生长素与油菜素甾醇相互作用机制的研究进展

植物生长发育是一个复杂、精细的调控过程,涉及细胞、组织和器官间多层次的信息交流,激素在其间发挥了重要调控作用．生长素和油菜素甾醇（BR）都能促进植物伸长,随着对其作用机制研究的深入,人们发现它们协同调控很多生理过程,对二者作用机制和信号转导的相互作用研究也成为激素研究领域的热点之一．对生长素和BR转导途径的揭示及直接下游基因的大规模发掘为二者通过相互作用调控不同生理过程的机制研究提供了重要线索．生长素和BR的相互作用涉及到下游基因转录的调控、信号组分互作以及合成代谢和极性运输等多层次的调控．     

Genetic approach towards the identification of auxin-cytokinin crosstalk components involved in root development

Phytohormones are important plant growth regulators that control many developmental processes, such as cell division, cell differentiation, organogenesis and morphogenesis. They regulate a multitude of apparently unrelated physiological processes, often with overlapping roles, and they mutually modulate their effects. These features imply important synergistic and antagonistic interactions between the various plant hormones. Auxin and cytokinin are central hormones involved in the regulation of plant growth and development, including processes determining root architecture, such as root pole establishment during early embryogenesis, root meristem maintenance and lateral root organogenesis. Thus, to control root development both pathways put special demands on the mechanisms that balance their activities and mediate their interactions. Here, we summarize recent knowledge on the role of auxin and cytokinin in the regulation of root architecture with special focus on lateral root organogenesis, discuss the latest findings on the molecular mechanisms of their interactions, and present forward genetic screen as a tool to identify novel molecular components of the auxin and cytokinin crosstalk.     

生长素在微生物调控根发育过程中的作用

很多微生物通过分泌生长素和生长素前体与植物建立了有益的关系并改变植物根系的形态结构,此外,微生物分泌的其他代谢产物也能改变植物生长素信号通路。因此,生长素和生长素信号通路在微生物调控植物根系发育的过程中起着至关重要的作用。该文从生长素合成、生长素信号和生长素极性运输3个方面总结了生长素在微生物调控植物根系发育过程中的作用,主要包括微生物增加了植物内源生长素的含量、增强了生长素的信号和调控PIN蛋白的表达水平,进而如何调控植物生理和分子水平来适应微生物对其根系的改变,为进一步开展该方面的研究奠定了基础。     

Genetic approach towards the identification of auxin–cytokinin crosstalk components involved in root development

Phytohormones are important plant growth regulators that control many developmental processes, such as cell division, cell differentiation, organogenesis and morphogenesis. They regulate a multitude of apparently unrelated physiological processes, often with overlapping roles, and they mutually modulate their effects. These features imply important synergistic and antagonistic interactions between the various plant hormones. Auxin and cytokinin are central hormones involved in the regulation of plant growth and development, including processes determining root architecture, such as root pole establishment during early embryogenesis, root meristem maintenance and lateral root organogenesis. Thus, to control root development both pathways put special demands on the mechanisms that balance their activities and mediate their interactions. Here, we summarize recent knowledge on the role of auxin and cytokinin in the regulation of root architecture with special focus on lateral root organogenesis, discuss the latest findings on the molecular mechanisms of their interactions, and present forward genetic screen as a tool to identify novel molecular components of the auxin and cytokinin crosstalk.     

Transmembrane auxin carrier systems – dynamic regulators of polar auxin transport

Recent investigations of the biochemistry, physiology and molecular genetics of polar auxin transport have greatly advanced our understanding of the process and of the part it plays in the regulation of development and in the responses of cells, tissues and organs to internal and external stimuli. The molecular and physiological characterization of mutants which exhibit lesions in polar auxin transport has led to the isolation and sequencing of genes which encode putative components of auxin carrier systems, or proteins which directly or indirectly regulate these systems. This work has revealed that specific auxin uptake and efflux carriers are coded not by single genes, but by whole families of genes, the expression of which is tissue or stimulus specific. Furthermore, evidence is accumulating rapidly that at least the auxin efflux carrier is a multi-component system consisting of both catalytic and regulatory subunits, including a separate phytotropin-binding protein. Other genes have been tentatively identified which code proteins that regulate the expression of genes coding auxin carrier components, or which regulate the intracellular traffic or activity of auxin carriers. Investigations of the turn-over and Golgi-mediated trafficking of auxin carrier proteins have revealed that essential components of at least the efflux carrier have a very short half-life in the plasma membrane and are replaced without the need for concurrent protein synthesis, leading to speculation that they might cycle between internal stores and the plasma membrane. The way is now clear for the development of specific molecular probes with which to investigate the intracellular transport and targeting of auxin carrier proteins.     

Phosphorylation of RACK1 in plants

Receptor for Activated C Kinase 1 (RACK1) is a versatile scaffold protein that interacts with a large, diverse group of proteins to regulate various signaling cascades. RACK1 has been shown to regulate hormonal signaling, stress responses and multiple processes of growth and development in plants. However, little is known about the molecular mechanism underlying these regulations. Recently, it has been demonstrated that Arabidopsis RACK1 is phosphorylated by an atypical serine/threonine protein kinase, WITH NO LYSINE 8 (WNK8). Furthermore, RACK1 phosphorylation by WNK8 negatively regulates RACK1 function by influencing its protein stability. These findings promote a new regulatory system in which the action of RACK1 is controlled by phosphorylation and subsequent protein degradation.     

Production and characterization of auxin-insensitive rice by overexpression of a mutagenized rice IAA protein

Since auxin was first isolated and characterized as a plant hormone, the underlying molecular mechanism of auxin signaling has been elucidated primarily in dicot plants represented by Arabidopsis. In monocot plants, the molecular mechanism of auxin signaling has remained unclear, despite various physiological experiments. To understand the function and mechanism of auxin signaling in rice ( Oryza sativa ), we focused on the IAA gene, a well-studied gene in Arabidopsis that serves as a negative regulator of auxin signaling. We found 24 IAA gene family members in the rice genome. OsIAA3 is one of these family members whose expression is rapidly increased in response to auxin. We produced transgenic rice harboring m OsIAA3 - GR , which can overproduce mutant OsIAA3 protein containing an amino acid change in domain II to cause a gain-of-function phenotype, by treatment with dexamethasone. The transgenic rice was insensitive to auxin and gravitropic stimuli, and exhibited short leaf blades, reduced crown root formation, and abnormal leaf formation. These results suggest that , in rice, auxin is important for development and its signaling is mediated by IAA genes.     

Polar auxin transport: controlling where and how much.

Auxin is transported through plant tissues, moving from cell to cell in a unique polar manner. Polar auxin transport controls important growth and developmental processes in higher plants. Recent studies have identified several proteins that mediate polar auxin transport and have shown that some of these proteins are asymmetrically localized, paving the way for studies of the mechanisms that regulate auxin transport. New data indicate that reversible protein phosphorylation can control the amount of auxin transport, whereas protein secretion through Golgi-derived vesicles and interactions with the actin cytoskeleton might regulate the localization of auxin efflux complexes.     

Genetic approaches to auxin action

Answers to long-standing questions concerning the molecular mechanism of auxin action and auxin's exact functions in plant growth and development are beginning to be uncovered through studies using mutant and transgenic plants. We review recent work in this area in vascular plants. A number of conclusions can be drawn from these studies. First, auxin appears essential for cell division and viability, as auxin auxotrophs isolated in tissue culture are dependent on auxin for growth and cannot be regenerated into plants even when auxin is supplied exogenously. Secondly, plants with transgenes that alter auxin levels are able to regulate cellular auxin concentrations by synthesis and conjugation; wild-type plants are probably also capable of such regulation. Thirdly, the phenotypes of transgenic plants with altered auxin levels and of mutant plants with altered sensitivity to auxin confirm earlier physiological studies which indicated a role for auxin in regulation of apical dominance, in development of roots and vascular tissue, and in the gravitropic response. Finally, the cloning of a mutationally identified gene important for auxin action, along with accumulating biochemical evidence, hints at a major role for protein degradation in the auxin response pathway.     

Genetic approaches to auxin action

Answers to long-standing questions concerning the molecular mechanism of auxin action and auxin's exact functions in plant growth and development are beginning to be uncovered through studies using mutant and transgenic plants. We review recent work in this area in vascular plants. A number of conclusions can be drawn from these studies. First, auxin appears essential for cell division and viability, as auxin auxotrophs isolated in tissue culture are dependent on auxin for growth and cannot be regenerated into plants even when auxin is supplied exogenously. Secondly, plants with transgenes that alter auxin levels are able to regulate cellular auxin concentrations by synthesis and conjugation; wild-type plants are probably also capable of such regulation. Thirdly, the phenotypes of transgenic plants with altered auxin levels and of mutant plants with altered sensitivity to auxin confirm earlier physiological studies which indicated a role for auxin in regulation of apical dominance, in development of roots and vascular tissue, and in the gravitropic response. Finally, the cloning of a mutationally identified gene important for auxin action, along with accumulating biochemical evidence, hints at a major role for protein degradation in the auxin response pathway.     

Molecular and cellular aspects of auxin-transport-mediated development

The plant hormone auxin is frequently observed to be asymmetrically distributed across adjacent cells during crucial stages of growth and development. These auxin gradients depend on polar transport and regulate a wide variety of processes, including embryogenesis, organogenesis, vascular tissue differentiation, root meristem maintenance and tropic growth. Auxin can mediate such a perplexing array of developmental processes by acting as a general trigger for the change in developmental program in cells where it accumulates and by providing vectorial information to the tissues by its polar intercellular flow. In recent years, a wealth of molecular data on the mechanism of auxin transport and its regulation has been generated, providing significant insights into the action of this versatile coordinative signal.     

Antagonistic regulation of PIN phosphorylation by PP2A and PINOID directs auxin flux

In plants, cell polarity and tissue patterning are connected by intercellular flow of the phytohormone auxin, whose directional signaling depends on polar subcellular localization of PIN auxin transport proteins. The mechanism of polar targeting of PINs or other cargos in plants is largely unidentified, with the PINOID kinase being the only known molecular component. Here, we identify PP2A phosphatase as an important regulator of PIN apical-basal targeting and auxin distribution. Genetic analysis, localization, and phosphorylation studies demonstrate that PP2A and PINOID both partially colocalize with PINs and act antagonistically on the phosphorylation state of their central hydrophilic loop, hence mediating PIN apical-basal polar targeting. Thus, in plants, polar sorting by the reversible phosphorylation of cargos allows for their conditional delivery to specific intracellular destinations. In the case of PIN proteins, this mechanism enables switches in the direction of intercellular auxin fluxes, which mediate differential growth, tissue patterning, and organogenesis.     

油菜素甾醇类与生长素的相互作用

油菜素甾醇类(brassinosteroid,BR)和生长素是两类重要的植物激素,二者在许多生理功能上存在相关性。近年来的研究表明,BR与生长素能协同调节基因表达,二者在代谢、运输和信号转导途径等不同层次上存在相互作用,并且这两种信号与其他信号转导途径,如激素信号转导途径和光信号转导途径之间也存在信号对话。现对BR与生长素之间这种复杂的相互作用进行评述。     

Auxin and cytokinin regulate each other's levels via a metabolic feedback loop

The hormones auxin and cytokinin are key regulators of plant growth and development. As they are active at minute concentrations and regulate dynamic processes, cell and tissue levels of the hormones are finely controlled developmentally, diurnally, and in response to environmental variables. This fine control, along with a regulation of the capacity to respond ensures that the appropriate type, duration and intensity of responses are elicited. We have recently discovered that cytokinin and auxin regulate the synthesis of each other, demonstrating a mechanism for mutual feed back and feed forward control of auxin and cytokinin levels. This regulatory loop could be important for many developmental processes in plants, i.e., in fine-tuning plant hormone levels in the developing meristems of the root and shoot apex. These findings could also give a molecular explanation for earlier observations of auxin and cytokinin effects on cell cultures,¹ where specific auxin and cytokinin ratios have been used to trigger different morphological events.Key words: auxin, cytokinin, biosynthesis, metabolism, signaling, root development, interactions     

Spermine-enhanced protein phosphorylation in human placenta

Polyamines are known to have a role in cell proliferation, differentiation, and protein synthesis. During pregnancy, major changes in polyamine levels occur in maternal serum, amniotic fluid, and placental tissue. Polyamine-activated phosphorylation has recently been proposed as a mechanism by which polyamines may regulate metabolic processes in target tissues. Polyamine-activated protein phosphorylation has not been studied in placenta. Homogenate membrane and cytosol fractions from human placenta were subjected to an endogenous protein phosphorylation assay using [gamma-32P]ATP in the presence and absence of the polyamines, spermine and spermidine, and the diamine, putrescine. Protein phosphorylation was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. When compared to basal levels, spermine (10(-3) M) significantly (P less than 0.001) stimulated 32P incorporation into phosphoproteins having molecular weights of 55,000 and 105,000. At this concentration spermidine and putrescine failed to stimulate phosphorylation. Half-maximal 32P incorporation was observed with 3.7 +/- 1.25 X 10(-4) M spermine. Polylysine enhanced the phosphorylation of phosphoproteins of the same molecular weight as those enhanced by spermine. Heparin and high Mg2+ inhibited spermine-induced phosphorylation. cAMP and Ca2+ did not stimulate phosphorylation of the spermine-dependent phosphoproteins. Spermine, however, acted as an antagonist for cAMP-dependent phosphorylation of a Mr 45,000 phosphoprotein.